Medroxyprogesterone derivatives from microbial transformation as anti-proliferative agents and acetylcholineterase inhibitors (combined in vitro and in silico approaches).

The fungal transformations of medroxyrogesterone (1) were investigated for the first time using Cunninghamella elegans, Trichothecium roseum, and Mucor plumbeus. The metabolites obtained are as following: 6ß, 20-dihydroxymedroxyprogesterone (2), 12ß-hydroxymedroxyprogesterone (3), 6ß, 11ß-dihydroxymedroxyprogesterone (4), 16ß-hydroxymedroxyprogesterone (5), 11α, 17-dihydroxy-6α-methylpregn-4-ene-3, 20-dione (6), 11-oxo-medroxyprogesterone (7), 6α-methyl-17α-hydroxypregn-1,4-diene-3,20-dione (8), and 6ß-hydroxymedroxyprogesterone (9), 15ß-hydroxymedroxyprogesterone (10), 6α-methyl-17α, 11ß-dihydroxy-5α-pregnan-3, 20-dione (11), 11ß-hydroxymedroxyprogesterone (12), and 11α, 20-dihydroxymedroxyprogesterone (13). Among all the microbial transformed products, the newly isolated biotransformed product 13 showed the most potent activity against proliferation of SH-SY5Y cells. Compounds 12, 5, 6, 9, 11, and 3 (in descending order of activity) also showed some extent of activity against SH-SY5Y tumour cell line. The never been reported biotransformed product, 2, showed the most potent inhibitory activity against acetylcholinesterase. Molecular modelling studies were carried out to understand the observed experimental activities, and also to obtain more information on the binding mode and the interactions between the biotransformed products, and enzyme.

Exploring sets of molecules from patents and relationships to other active compounds in chemical space networks.

Patents from medicinal chemistry represent a rich source of novel compounds and activity data that appear only infrequently in the scientific literature. Moreover, patent information provides a primary focal point for drug discovery. Accordingly, text mining and image extraction approaches have become hot topics in patent analysis and repositories of patent data are being established. In this work, we have generated network representations using alternative similarity measures to systematically compare molecules from patents with other bioactive compounds, visualize similarity relationships, explore the chemical neighbourhood of patent molecules, and identify closely related compounds with different activities. The design of network representations that combine patent molecules and other bioactive compounds and view patent information in the context of current bioactive chemical space aids in the analysis of patents and further extends the use of molecular networks to explore structure-activity relationships.

Probing the action of a novel anti-leukaemic drug therapy at the single cell level using modern vibrational spectroscopy techniques.

Acute myeloid leukaemia (AML) is a life threatening cancer for which there is an urgent clinical need for novel therapeutic approaches. A redeployed drug combination of bezafibrate and medroxyprogesterone acetate (BaP) has shown anti-leukaemic activity in vitro and in vivo. Elucidation of the BaP mechanism of action is required in order to understand how to maximise the clinical benefit. Attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, Synchrotron radiation FTIR (S-FTIR) and Raman microspectroscopy are powerful complementary techniques which were employed to probe the biochemical composition of two AML cell lines in the presence and absence of BaP. Analysis was performed on single living cells along with dehydrated and fixed cells to provide a large and detailed data set. A consideration of the main spectral differences in conjunction with multivariate statistical analysis reveals a significant change to the cellular lipid composition with drug treatment; furthermore, this response is not caused by cell apoptosis. No change to the DNA of either cell line was observed suggesting this combination therapy primarily targets lipid biosynthesis or effects bioactive lipids that activate specific signalling pathways.

[Biodehydrogenation of 11beta-hydroxyl melroxyprogesterone by Arthrobacter simplex UR016 in microemulsion system].

To improve mass transfer and enhance the yield for C(1,2) biodehydrogenation of steroid 11beta-hydroxyl medroxyprogesterone, we carried out the dehydrogenation reaction of 11beta-hydroxyl medroxyprogesterone in an oil-in-water (O/W) microemulsion by Arthrobacter simplex UR016. We studied the effects of system composition, dehydrogenation temperature and substrate concentration on microbial transformation. We formulated a suitable O/W microemulsion system with Arthrobacter simplex UR016 culture broth as aqueous phase, 10 g/L of edible oil as oil phase, 4 g/L of Tween-O80 and 7% (V/V) alcohol as surfactant and cosurfactant. The optimal dehydrogenation temperature was 33 degrees C. The results showed that in Tween-80/alcohol/edible oil/water microemulsion system, the hydrophobic steroid was solubilised and diffused effectively, with the maximum conversion rate of 88.6% at 46 h under 4 g/L substrate concentration, an increase of 66.2% compared to that in aqueous system. The C(1,2) biodehydrogenation of 11beta-hydroxyl medroxyprogesterone is more efficient in water-edible oil microemulsion system than in aqueous system.

Ozone oxidation of pharmaceuticals, endocrine disruptors and pesticides during drinking water treatment.

This study investigates the oxidation of pharmaceuticals, endocrine disrupting compounds and pesticides during ozonation applied in drinking water treatment. In the first step, second-order rate constants for the reactions of selected compounds with molecular ozone (k(O3)) were determined in bench-scale experiments at pH 8.10: caffeine (650+/-22M(-1)s(-1)), progesterone (601+/-9M(-1)s(-1)), medroxyprogesterone (558+/-9M(-1)s(-1)), norethindrone (2215+/-76M(-1)s(-1)) and levonorgestrel (1427+/-62M(-1)s(-1)). Compared to phenolic estrogens (estrone, 17beta-estradiol, estriol and 17alpha-ethinylestradiol), the selected progestogen endocrine disruptors reacted far slower with ozone. In the second part of the study, bench-scale experiments were conducted with surface waters spiked with 16 target compounds to assess their oxidative removal using ozone and determine if bench-scale results would accurately predict full-scale removal data. Overall, the data provided evidence that ozone is effective for removing trace organic contaminants from water with ozone doses typically applied in drinking water treatment. Ozonation removed over 80% of caffeine, pharmaceuticals and endocrine disruptors within the CT value of about 2 mg min L(-1). As expected, pesticides were found to be the most recalcitrant compounds to oxidize. Caffeine can be used as an indicator compound to gauge the efficacy of ozone treatment.

Not all progestins are the same: implications for usage.

Recent clinical evidence showing unexpected side-effects of progestins used in contraception and hormone replacement therapy has highlighted the importance of choice of synthetic progestin. The molecular mechanisms of action of the relatively nonspecific and most widely used synthetic progestins, medroxyprogesterone acetate and norethisterone, are discussed in the context of this recent clinical evidence. Future directions involving a more mechanism-based approach for improved therapeutics, with greater specificity and fewer side-effects, are discussed.

A chemiluminescence enzyme immunoassay (CLEIA) for the determination of medroxyprogesterone acetate in human serum.

An enzyme immunoassay with chemiluminescence detection (CLEIA) for measuring serum levels of medroxyprogesterone acetate (MPA), a synthetic progestational agent currently used in fertility control and hormonal cancer, is reported. The polyclonal antiserum was obtained by immunizing rabbits with the synthetized 17-hemisuccinate derivative of medroxyprogesterone (MPS) coupled to serum albumin. This antiserum does not display any cross reactivity with extracted metabolites or with corticosteroid analogs with modifications at positions 11 and 16. The same MPS coupled to alkaline phosphatase is used as tracer. For the chemiluminescent detection system, adamantyl-1,2-dioxetane phosphate is selected as substrate. The typical standard curve ranges from 18.5 to 1182 pg per well and displays a slope factor of 0.74, with an ED50 of 143.8 pg of MPA per well and a minimum detectable and maximal level of 0.83 and 12,400 pg per well respectively. The assay has been validated on spiked serum samples in terms of precision (intra- and interassay coefficient variations of less than 8% and 13%, respectively), and of accuracy (mean recovery 105%). The validation on clinical samples demonstrates a good correlation of MPA serum values obtained both by radioimmunoassay and CLEIA. This specific and sensitive CLEIA, which requires less than 100 microliters of serum sample, appears to be an interesting alternative for the monitoring of serum levels of MPA in humans.